Tet-on circRNA Expression Vector vector (V000415)

Basic Vector Information

      • Vector Name:
      • Tet-on circRNA Expression Vector
      • Antibiotic Resistance:
      • Low Ampicillin
      • Length:
      • 10054 bp
      • Type:
      • Mammalian Expression, Lentiviral
      • Copy Number:
      • High Copy
      • Promoter:
      • pTet
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • none specified

Tet-on circRNA Expression Vector vector Vector Map

Tet-on circRNA Expression Vector10054 bp500100015002000250030003500400045005000550060006500700075008000850090009500100003' LTRHIV-1 PsiRREgp41 peptidecPPT/CTStight TRE promoterKozak sequenceEGFP-NmCherry-FIRESEXFP-REGFP-CWPRE3' LTR (Delta-U3)pBRforEcoAmpR promoterAmpRoriL4440SV40pro-FCAP binding sitelac promoterlac operatorM13 revEBV-rev

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

Tet-on circRNA Expression Vector vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               10054 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  The vector can ef_ciently produce a circRNA (circmCherry) from a 
            split EGFP gene after Dox treatment..
ACCESSION   .
VERSION     .
KEYWORDS    Tet-on circRNA Expression Vector
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10054)
  AUTHORS   Zhang Y, Xue W, Li X, Zhang J, Chen S, Zhang JL, Yang L, Chen LL
  TITLE     The Biogenesis of Nascent Circular RNAs.
  JOURNAL   Cell Rep. 2016 Apr 19;15(3):611-24. doi: 
            10.1016/j.celrep.2016.03.058. Epub 2016 Apr 7.
  PUBMED    27068474
REFERENCE   2  (bases 1 to 10054)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2016.03.058"; journalName: "Cell Rep"; date: 
            "2016-04-19- 19"; volume: "15"; issue: "3"; pages: "611-24"
FEATURES             Location/Qualifiers
     source          1..10054
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2228..2541
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2235..2253
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2271..2289
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2306..2324
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2342..2360
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2378..2396
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2413..2431
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2449..2467
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2514..2538
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2569..2578
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(2620..2641)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3942..3961
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4289..4857
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4307..4859
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4451..4468)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4678..4697
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(5890..5909)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6237..6258
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6444..7032
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6497..6517)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6915..6926)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7107..7340
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7459..7477)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7545..7649
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7650..8510
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7868..7887)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8681..9269
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9170..9189
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9423..9440
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     9532..9551
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    9680..9701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9716..9746
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9754..9770
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     9759..9781
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9778..9794
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9778..9794
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9844..9863
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"

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